CN115044580A - Compound microbial agent for removing sulfide and preparation method thereof - Google Patents
Compound microbial agent for removing sulfide and preparation method thereof Download PDFInfo
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- CN115044580A CN115044580A CN202210924944.2A CN202210924944A CN115044580A CN 115044580 A CN115044580 A CN 115044580A CN 202210924944 A CN202210924944 A CN 202210924944A CN 115044580 A CN115044580 A CN 115044580A
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- compound microbial
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- sulfide
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- 150000001875 compounds Chemical class 0.000 title claims abstract description 41
- 230000000813 microbial effect Effects 0.000 title claims abstract description 37
- UCKMPCXJQFINFW-UHFFFAOYSA-N Sulphide Chemical compound [S-2] UCKMPCXJQFINFW-UHFFFAOYSA-N 0.000 title claims abstract description 35
- 238000002360 preparation method Methods 0.000 title claims abstract description 17
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 36
- 239000010410 layer Substances 0.000 claims abstract description 29
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 25
- 239000012790 adhesive layer Substances 0.000 claims abstract description 14
- 238000005538 encapsulation Methods 0.000 claims abstract description 11
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 45
- 241000607598 Vibrio Species 0.000 claims description 22
- 235000011187 glycerol Nutrition 0.000 claims description 17
- 239000001963 growth medium Substances 0.000 claims description 17
- 239000000843 powder Substances 0.000 claims description 17
- 150000004763 sulfides Chemical class 0.000 claims description 14
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 12
- 241000589516 Pseudomonas Species 0.000 claims description 12
- 239000000084 colloidal system Substances 0.000 claims description 12
- 238000011081 inoculation Methods 0.000 claims description 12
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 11
- 238000000034 method Methods 0.000 claims description 10
- 210000003097 mucus Anatomy 0.000 claims description 9
- 239000002245 particle Substances 0.000 claims description 9
- 235000013339 cereals Nutrition 0.000 claims description 8
- 239000002068 microbial inoculum Substances 0.000 claims description 8
- 239000013049 sediment Substances 0.000 claims description 8
- 238000006477 desulfuration reaction Methods 0.000 claims description 7
- 230000023556 desulfurization Effects 0.000 claims description 7
- 230000001580 bacterial effect Effects 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 6
- 238000012216 screening Methods 0.000 claims description 6
- 239000000725 suspension Substances 0.000 claims description 6
- 241000894006 Bacteria Species 0.000 claims description 4
- 239000013535 sea water Substances 0.000 claims description 4
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 claims description 3
- 229920002472 Starch Polymers 0.000 claims description 3
- 235000019270 ammonium chloride Nutrition 0.000 claims description 3
- 239000011248 coating agent Substances 0.000 claims description 3
- 238000000576 coating method Methods 0.000 claims description 3
- VDQVEACBQKUUSU-UHFFFAOYSA-M disodium;sulfanide Chemical compound [Na+].[Na+].[SH-] VDQVEACBQKUUSU-UHFFFAOYSA-M 0.000 claims description 3
- 229960002413 ferric citrate Drugs 0.000 claims description 3
- 235000013312 flour Nutrition 0.000 claims description 3
- 229910017053 inorganic salt Inorganic materials 0.000 claims description 3
- NPFOYSMITVOQOS-UHFFFAOYSA-K iron(III) citrate Chemical compound [Fe+3].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NPFOYSMITVOQOS-UHFFFAOYSA-K 0.000 claims description 3
- 239000002609 medium Substances 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 238000004806 packaging method and process Methods 0.000 claims description 3
- 238000005096 rolling process Methods 0.000 claims description 3
- 229910052979 sodium sulfide Inorganic materials 0.000 claims description 3
- 235000019698 starch Nutrition 0.000 claims description 3
- 239000008107 starch Substances 0.000 claims description 3
- 241000605716 Desulfovibrio Species 0.000 claims description 2
- 230000000694 effects Effects 0.000 abstract description 7
- 239000003124 biologic agent Substances 0.000 abstract description 3
- 230000005484 gravity Effects 0.000 abstract description 3
- 239000000463 material Substances 0.000 abstract description 3
- 150000003568 thioethers Chemical class 0.000 abstract 3
- 239000003643 water by type Substances 0.000 description 8
- 239000003610 charcoal Substances 0.000 description 6
- 241000251468 Actinopterygii Species 0.000 description 4
- 239000000969 carrier Substances 0.000 description 4
- 238000009364 mariculture Methods 0.000 description 4
- 241000191938 Micrococcus luteus Species 0.000 description 3
- 241000589540 Pseudomonas fluorescens Species 0.000 description 3
- 241000588626 Acinetobacter baumannii Species 0.000 description 2
- 241000186063 Arthrobacter Species 0.000 description 2
- 241000194032 Enterococcus faecalis Species 0.000 description 2
- 241000194105 Paenibacillus polymyxa Species 0.000 description 2
- 241000187563 Rhodococcus ruber Species 0.000 description 2
- 230000032683 aging Effects 0.000 description 2
- 229940032049 enterococcus faecalis Drugs 0.000 description 2
- 231100000252 nontoxic Toxicity 0.000 description 2
- 230000003000 nontoxic effect Effects 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 241000589291 Acinetobacter Species 0.000 description 1
- 241001524201 Arthrobacter crystallopoietes Species 0.000 description 1
- 241000179039 Paenibacillus Species 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 239000010903 husk Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000005416 organic matter Substances 0.000 description 1
- 230000033116 oxidation-reduction process Effects 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000029058 respiratory gaseous exchange Effects 0.000 description 1
- 238000004062 sedimentation Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/10—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a carbohydrate
-
- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/36—Adaptation or attenuation of cells
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/38—Chemical stimulation of growth or activity by addition of chemical compounds which are not essential growth factors; Stimulation of growth by removal of a chemical compound
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/04—Enzymes or microbial cells immobilised on or in an organic carrier entrapped within the carrier, e.g. gel or hollow fibres
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/14—Enzymes or microbial cells immobilised on or in an inorganic carrier
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
- C02F2103/08—Seawater, e.g. for desalination
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- C—CHEMISTRY; METALLURGY
- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F2103/00—Nature of the water, waste water, sewage or sludge to be treated
- C02F2103/20—Nature of the water, waste water, sewage or sludge to be treated from animal husbandry
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- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
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- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/38—Pseudomonas
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- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/63—Vibrio
Abstract
The invention belongs to the technical field of microbial agents, and particularly relates to a compound microbial agent for removing sulfides and a preparation method thereof, wherein the compound microbial agent comprises a carrier, an attachment layer and an encapsulation layer which are arranged from inside to outside; the adhesive layer is attached to the carrier and then encapsulated inside by an encapsulation layer; the overall average density of the compound microbial agent is greater than the density of a water area where the sulfide is located. According to the compound microbial agent for removing sulfides and the preparation method thereof, the carrier is arranged, so that the biological agent can be smoothly brought to the bottom of a water area under the action of the self gravity of the carrier, the effect of removing sulfides from the bottom of the water area is further realized, and the problem of secondary pollution is avoided due to the adoption of a completely pollution-free material.
Description
Technical Field
The invention relates to the technical field of microbial agents, in particular to a compound microbial agent for removing sulfides and a preparation method thereof.
Background
China is a large mariculture country, and in offshore culture areas, people often add a large amount of organic baits to the culture areas in order to improve the yield of mariculture, so that organic matter pollution of water bodies in the culture areas is serious. In the sediment-water interface environment of the offshore culture area, the aerobic degradation of organic matters causes the large consumption of dissolved oxygen in the water body, and the oxidation-reduction potential (Eh) of the water body is reduced. Under the condition, Sulfate-reducing bacteria (SRB) in the sediments generate a large amount of sulfides through a Sulfate breathing process, the sulfides are accumulated in a sediment-water interface in a large amount and are diffused to an upper water body, the toxic action of the high-concentration sulfides seriously threatens seawater, especially benthic organisms, so that the aging phenomenon of the sediments in a culture area is caused, and the healthy development of the mariculture industry is influenced.
At present, aiming at the problem of 'aging' phenomenon of a mariculture area, particularly the problem of overhigh concentration of sulfide, people mostly adopt a physical or chemical method for treatment, particularly at the deep water bottom, most carriers of the microbial inoculum are powdery and light in weight, most carriers are consumed after being used for the carriers and mixed with water along with precipitation, and on the other hand, the adopted carriers are easy to generate secondary pollution.
Disclosure of Invention
The invention provides a compound microbial agent for removing sulfide and a preparation method thereof based on the technical problems of the existing compound microbial agent for removing sulfide.
The invention provides a compound microbial agent for removing sulfides, which comprises a carrier, an attachment layer and an encapsulation layer, wherein the carrier, the attachment layer and the encapsulation layer are arranged from inside to outside;
the adhesive layer is attached to the carrier and then encapsulated inside by an encapsulation layer;
the overall average density of the compound microbial agent is greater than the density of a water area where the sulfide is located.
Preferably, the carrier comprises powders including but not limited to carbon powder, starch and flour mixed in any proportion.
Through the technical scheme, the carbon powder can adopt the existing charcoal powder, the charcoal not only has the function of purifying water quality, but also can not cause secondary pollution, and the charcoal has low cost, is easy to obtain and is convenient to operate.
Preferably, the carrier also comprises colloid with density larger than water density, and the colloid is mixed with the powder to be made into particles with diameter of 3-10 mm.
Through the technical scheme, the density of the carrier is greater than the water density in the treated water area, and rapid sedimentation can be facilitated.
Preferably, the adhesion layer comprises any one or more of bacillus, vibrio, pseudomonas and devulcanization vibrio for removing sulfide in any ratio.
Preferably, the bacillus, vibrio, pseudomonas and devulcanizing vibrio are mixed with the powder crushed from the grain husk, and then are added with colloid to form mucus, and then are wrapped and attached on the outer surface of the carrier.
Through the technical scheme, the cereal shell can better have the removing effect of the microbial inoculum on sulfides, and is environment-friendly and energy-saving.
Preferably, the packaging layer includes, but is not limited to, glycerin, and the glycerin includes one or more of natural glycerin and synthetic glycerin mixed in any proportion.
By the technical scheme, the glycerol is completely nontoxic, and even if the glycerol is eaten by fish by mistake, the species in the water area can not be damaged artificially.
A preparation method of a compound microbial agent for removing sulfides comprises the following steps:
s1, preparing the carrier into particles, and then coating the adhesive layer with the adsorbed and removed sulfide on the outer surface of the carrier;
s2, screening bacillus, vibrio, pseudomonas and desulfurization vibrio in the adhesion layer, mixing the screened bacillus, vibrio, pseudomonas and desulfurization vibrio with the powder crushed from the grain hulls, adding colloid to form mucus, and wrapping and adhering the mucus on the outer surface of the carrier;
s3, smearing the glycerol on the adhesive layer, putting the adhesive layer into carbon powder, and rolling and wrapping the carbon powder into particles which can be held by hands for use.
Preferably, the screening method in step S2 is:
p1, collecting surface sediments in the offshore culture area as an inoculation source to prepare bacterial suspension;
p2, inoculating the bacterial suspension obtained in the step P1 to a primary selection culture medium with sulfide concentration of 600mg/l in proportion, and performing acclimation culture to obtain a primary culture solution;
p3, inoculating the primary culture solution for inoculation in the step P2 into a secondary selection culture medium with the sulfide concentration of 800mg/l according to a proportion, and obtaining a secondary culture solution after acclimatization culture;
p4, inoculating the secondary culture solution for inoculation in the step P3 into a tertiary selection culture medium with the sulfide concentration of 1000mg/l according to a proportion, and obtaining a tertiary culture solution after acclimatization culture;
and P5, inoculating the third-stage culture solution for inoculation in the step P4 into an inorganic salt culture medium in proportion, and obtaining the compound microbial inoculum for removing sulfide in sediments after bacteria grow to a logarithmic phase.
Preferably, the composition of the primary selection medium in step P2 is as follows:
k2HPO 4: 1.2 g/L, KH2PO 4: 1.2 g/L, NH4Cl: 0.4 g/L, MgCl 2: 0.2g/L, ferric citrate: 0.01g/L, NaHCO 3: 2g/L, Na 2S: 1.5 g/L.
Through the technical scheme, more preparation raw materials of the culture medium can be obtained, and the diversity is increased.
Preferably, the Na2S concentration in the secondary and tertiary culture solution re-selection culture medium compositions in the steps P3 and P4 is respectively increased to 2.0g/L and 2.5g/L, and the rest is unchanged;
then, the preparation is carried out by using sterilized aged seawater.
Through above-mentioned technical scheme, the preparation culture solution that can become more meticulous facilitates the use.
The beneficial effects of the invention are as follows:
1. through setting up the carrier, can realize that biological agent is brought into the bottom in waters smoothly under the self action of gravity of carrier, and then realize beginning the effect of getting rid of the sulphide from the waters bottom, and adopt the material that does not have the pollution completely, can not cause secondary pollution's problem.
2. Through setting up the adhesion layer, can be fine be connected with the carrier with compound microbial inoculum, conveniently be brought into by the carrier and deposit to the waters bottom.
3. Through setting up the encapsulation layer, can make the protection before using with the adhesion layer, also can prevent simultaneously by the fish biology in the waters swallow in a large number and become invalid.
4. By the adoption of the preparation method, the compound microbial agent can be conveniently and quickly prepared, and the problem that the compound microbial agent fails or is low in effect due to errors is solved.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments.
Example one
The compound microbial agent for removing the sulfide comprises a carrier, an attachment layer and an encapsulation layer which are arranged from inside to outside.
Further, the carrier comprises powder, and the powder comprises but is not limited to carbon powder, starch and flour which are mixed according to any proportion. The carbon powder can be the existing charcoal powder, the charcoal not only has the function of purifying water quality, but also can not cause secondary pollution, and the charcoal powder is low in cost, easy to obtain and convenient to operate.
Further, the carrier also comprises colloid with the density larger than that of water, and the colloid and the powder are mixed to be made into particles with the diameter of 3-10 mm. The density of the carrier is greater than the density of water in the treated water area, which can facilitate rapid settling down.
Through setting up the carrier, can realize that biological agent is brought into the bottom in waters smoothly under the self action of gravity of carrier, and then realize beginning the effect of getting rid of the sulphide from the waters bottom, and adopt the material that does not have the pollution completely, can not cause secondary pollution's problem.
And after the adhesive layer is attached to the carrier, the adhesive layer is encapsulated inside the carrier by using an encapsulation layer.
The overall average density of the compound microbial agent is greater than the density of a water area where the sulfide is located.
Further, the attachment layer comprises any one or combination of more of bacillus, vibrio, pseudomonas and devulcanization vibrio in any proportion for removing sulfide.
Further, the bacillus, the vibrio, the pseudomonas and the desulfovibrio are mixed with the powder crushed from the grain hulls, and then colloid is added to form mucus which is wrapped and attached on the outer surface of the carrier. The cereal shell can have the effect of removing sulfide by microbial inoculum, and is environment-friendly and energy-saving.
Through setting up the adhesion layer, can be fine be connected with the carrier with compound microbial inoculum, conveniently be brought into by the carrier and deposit to the waters bottom.
Further, the packaging layer includes, but is not limited to, glycerin, and the glycerin includes one or more of natural glycerin and synthetic glycerin mixed in any proportion.
The glycerol is completely nontoxic, and even if the glycerol is eaten by fish by mistake, the glycerol can not cause artificial damage to the species in the water area.
Through setting up the encapsulation layer, can make the protection before using with the adhesion layer, also can prevent simultaneously by the fish biology in the waters swallow in a large number and become invalid.
Example two
A preparation method of a compound microbial agent for removing sulfides comprises the following steps:
s1, preparing the carrier into particles, and then coating the adhesive layer with the adsorbed and removed sulfide on the outer surface of the carrier;
s2, screening bacillus, vibrio, pseudomonas and desulfurization vibrio in the adhesion layer, mixing the screened bacillus, vibrio, pseudomonas and desulfurization vibrio with the powder crushed from the grain hulls, adding colloid to form mucus, and wrapping and adhering the mucus on the outer surface of the carrier;
s3, smearing the glycerol on the adhesive layer, putting the adhesive layer into carbon powder, rolling and wrapping the carbon powder into particles which can be held by hands for use.
Further, the screening method in step S2 is:
p1, collecting surface sediments in the offshore culture area as an inoculation source to prepare bacterial suspension;
p2, inoculating the bacterial suspension obtained in the step P1 to a primary selection culture medium with sulfide concentration of 600mg/l in proportion, and performing acclimation culture to obtain a primary culture solution;
p3, inoculating the primary culture solution for inoculation in the step P2 into a secondary selection culture medium with the sulfide concentration of 800mg/l according to a proportion, and obtaining a secondary culture solution after acclimatization culture;
p4, inoculating the secondary culture solution for inoculation in the step P3 into a tertiary selection culture medium with the sulfide concentration of 1000mg/l according to a proportion, and obtaining a tertiary culture solution after acclimatization culture;
and P5, inoculating the third-stage culture solution for inoculation in the step P4 into an inorganic salt culture medium in proportion, and obtaining the compound microbial inoculum for removing sulfide in sediments after bacteria grow to a logarithmic phase.
Further, the composition of the primary selection medium in step P2 is as follows:
k2HPO 4: 1.2 g/L, KH2PO 4: 1.2 g/L, NH4Cl: 0.4 g/L, MgCl 2: 0.2g/L, ferric citrate: 0.01g/L, NaHCO 3: 2g/L, Na 2S: 1.5 g/L.
Can obtain more preparation raw materials of the culture medium and increase the diversity.
Further, the Na2S concentration in the secondary and tertiary culture solution re-selection culture medium compositions in the steps P3 and P4 is respectively increased to 2.0g/L and 2.5g/L, and the others are unchanged; then, the preparation is carried out by using sterilized aged seawater. Can finely produce the culture solution, and is convenient to use.
The compound microorganism strain can also be Rhodococcus ruber (Rhodococcus ruber) ATCC 15906; the Micrococcus luteus is Micrococcus luteus (Micrococcus luteus) ATCC 49442; the enterococcus faecalis is enterococcus faecalis (E nterococcus faecalis) ATCC 29212; said Acinetobacter baumannii is Acinetobacter baumannii (Acinetobacter erbaumanii) ATCC 19606; the Arthrobacter crystallopoiicus is Arthrobacter crystallopoiensis (Arthrobacter crystallopoietes) ATCC 15481; the paenibacillus polymyxa is paenibacillus polymyxa (paenibacillus ly myxa) ATCC 842; the pseudomonas fluorescens is any combination of multiple pseudomonas fluorescens (P. fluorescences) ATCC49642, and is prepared by adopting the preparation method.
By the adoption of the preparation method, the compound microbial agent can be conveniently and quickly prepared, and the problem that the compound microbial agent fails or is low in effect due to errors is solved.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.
Claims (10)
1. A compound microbial agent for removing sulfides is characterized in that: the compound microbial agent comprises a carrier, an attachment layer and an encapsulation layer which are arranged from inside to outside;
the adhesive layer is attached to the carrier and then encapsulated inside by an encapsulation layer;
the overall average density of the compound microbial agent is greater than the density of a water area where the sulfide is located.
2. The compound microbial agent for removing sulfides according to claim 1, wherein: the carrier comprises powder, and the powder comprises but is not limited to carbon powder, starch and flour which are mixed according to any proportion.
3. The compound microbial agent for removing sulfides according to claim 2, wherein: the carrier also comprises colloid with the density larger than that of water, and the colloid and the powder are mixed to be made into particles with the diameter of 3-10 mm.
4. The compound microbial agent for removing sulfides according to claim 3, wherein: the attachment layer comprises any one or combination of more of bacillus, vibrio, pseudomonas and desulfurization vibrio in any proportion for removing sulfide.
5. The compound microbial agent for removing sulfides according to claim 4, wherein: the bacillus, the vibrio, the pseudomonas and the desulfovibrio are mixed with the powder crushed from the grain hulls, and then colloid is added to form mucus which is coated and attached on the outer surface of the carrier.
6. The compound microbial agent for removing sulfides according to claim 5, wherein: the packaging layer comprises but is not limited to glycerin, and the glycerin comprises any one or more of natural glycerin and synthetic glycerin mixed in any proportion.
7. The method for preparing the compound microbial agent for removing the sulfide as claimed in claim 6, wherein the compound microbial agent comprises: the method comprises the following steps:
s1, preparing the carrier into particles, and then coating the adhesive layer with the adsorbed and removed sulfide on the outer surface of the carrier;
s2, screening bacillus, vibrio, pseudomonas and desulfurization vibrio in the adhesion layer, mixing the screened bacillus, vibrio, pseudomonas and desulfurization vibrio with the powder crushed from the grain hulls, adding colloid to form mucus, and wrapping and adhering the mucus on the outer surface of the carrier;
s3, smearing the glycerol on the adhesive layer, putting the adhesive layer into carbon powder, and rolling and wrapping the carbon powder into particles which can be held by hands for use.
8. The method for preparing the compound microbial agent for removing the sulfide as claimed in claim 7, wherein the compound microbial agent comprises: the screening method in step S2 is:
p1, collecting surface sediments in the offshore culture area as an inoculation source to prepare bacterial suspension;
p2, inoculating the bacterial suspension obtained in the step P1 to a primary selection culture medium with sulfide concentration of 600mg/l in proportion, and performing acclimation culture to obtain a primary culture solution;
p3, inoculating the primary culture solution for inoculation in the step P2 into a secondary selection culture medium with the sulfide concentration of 800mg/l according to a proportion, and obtaining a secondary culture solution after acclimatization culture;
p4, inoculating the secondary culture solution for inoculation in the step P3 into a tertiary selection culture medium with the sulfide concentration of 1000mg/l according to a proportion, and obtaining a tertiary culture solution after acclimatization culture;
and P5, inoculating the third-stage culture solution for inoculation in the step P4 into an inorganic salt culture medium in proportion, and obtaining the compound microbial inoculum for removing sulfide in sediments after bacteria grow to a logarithmic phase.
9. The method for preparing the compound microbial agent for removing the sulfide according to claim 8, wherein the compound microbial agent comprises the following components in percentage by weight: the composition of the primary selection medium in step P2 is as follows:
k2HPO 4: 1.2 g/L, KH2PO 4: 1.2 g/L, NH4Cl: 0.4 g/L, MgCl 2: 0.2g/L, ferric citrate: 0.01g/L, NaHCO 3: 2g/L, Na 2S: 1.5 g/L.
10. The method for preparing the compound microbial agent for removing the sulfide as claimed in claim 9, wherein the compound microbial agent comprises: the Na2S concentration in the secondary and tertiary culture solution re-selection culture medium compositions in the steps P3 and P4 is respectively increased to 2.0g/L and 2.5g/L, and the others are unchanged;
then, the preparation is carried out by using sterilized aged seawater.
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